The generation of antibody combining sites containing catalytic residues

Ciba Found Symp. 1991:159:118-28; discussion 128-34. doi: 10.1002/9780470514108.ch9.

Abstract

To expand the scope of antibody-catalysed reactions to those involving rate-limiting proton abstraction, such as elimination, isomerization and condensation reactions, we developed a new strategy--hapten charge complementarity. A hapten containing a benzyl ammonium group was used to elicit a specific base, a carboxylate, in the combining site of an antibody that catalysed a beta-elimination reaction. This was the first example of the use of a hapten to elicit a specific catalytic residue in an antibody combining site. A variety of kinetic and chemical modification experiments strongly suggest that a specific Asp or Glu residue in the combining site is responsible for catalysis. Preliminary results indicate that in addition to charge-charge complementarity, the nucleophilic reactivity of amino acid residues (Ser, Thr, Lys, Asp, Glu, Cys) in antibodies can be used as a selection tool. Antibodies were raised against a reactive epoxide group to elicit an antibody containing a uniquely reactive carboxylate or thiol group. Antibodies which bind the epoxide do catalyse a beta-elimination reaction, indicating the presence of a specific base in the combining site. Antibodies elicited to two closely related haptens do not catalyse the beta-elimination reaction.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies / metabolism*
  • Antigens / immunology
  • Binding Sites, Antibody / physiology*
  • Catalysis
  • Electrochemistry
  • Haptens / immunology

Substances

  • Antibodies
  • Antigens
  • Haptens